Glycerol oxidase and process for the production thereof and method for the quantitative determination of glycerol by glycerol oxidase

Abstract

Glycerol oxidase is an oxidizing enzyme of glycerol characterized by its ability to oxidize glycerol in the presence of oxygen to form hydrogen peroxide and glyceraldehyde. This enzyme is produced by cultivation of a microorganism belonging to the genus Aspergillus or the genus Neurospora in a nutrient medium.

What is claimed is: 
     
       1. A method for the quantitative determination of glycerol in solution which comprises reacting glycerol oxidase, an enzyme having the ability to oxidize glycerol in the presence of oxygen to form hydrogen peroxide and gylceraldehyde, with the glycerol in the presence of oxygen and measuring the amount of oxygen consumed, or the amount of hydrogen peroxide or glyceraldehyde formed in an aqueous medium by the action of said glycerol oxidase, said amount being an indication of the concentration of glycerol in said solution. 
     
     
       2. The method of claim 1, wherein the formed hydrogen peroxide is quantitatively determined by reacting the peroxide with 4-aminoantipyrine and phenol in the presence of peroxidase to obtain quinoneimine pigment, which is quantitatively determined by measuring the optical density of the reaction solution at 500 nm. 
     
     
       3. The method of claim 1, wherein the glyceraldehyde formed is reacted with 2,4-dinitrophenylhydrazine, to form 2,4-dinitrophenylhydrazone and the 2,4-dinitrophenylhydrazone is quantitatively determined by colorimetry, this providing a quantitative determination of glycerol. 
     
     
       4. The method of claim 1, wherein the amount of oxygen consumed is measured by an oxygen electrode and a Warburg's manometer.